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Multiple myeloma is characterized by malignant cells which produce high amounts of monoclonal immunoglobulin. Myeloma cells are, therefore, dependent on effective protein degradation. Proteasomal protein degradation is targeted by proteasome inhibitors in routine care. Autophagic protein degradation is currently not targeted in myeloma treatment. This Phase I trial showed that the combination of the proteasome inhibitor carfilzomib and the autophagy inhibitor hydroxychloroquine was well tolerated in patients with relapsed/refractory multiple myeloma. Adverse events were mostly Grades 1 and 2. An overall response rate of 44% indicates a meaningful clinical efficacy of this combination. Trial Registration : The study was registered at clinicaltrials.gov # NCT04163107., Competing Interests: Tobias S. Slørdahl has received honoraria from Takeda, Celgene, Amgen, Janssen, AbbVie, Pfizer; consultancy for Bristol Myers Squibb, GSK, Pfizer, Menarini Group, Sanofi; advisory board consultancy: Bristol Myers Squibb, Sanofi, Amgen, Celgene, GSK, Janssen. Frida Bugge Askeland has received consulting fees from Janssen, Sanofi, and Amgen, honoraria/payment from Janssen and Sanofi, participated in advisory boards for Janssen and Sanofi, and received research support from Sanofi. Fredrik Schjesvold has received grants from Targovax, consulting fees from GSK, BMS, Oncopeptides, XNK Therapeutics, Takeda, Janssen, Sanofi, and Galapagos, honoraria/payment from Amgen, BMS, Takeda, Sanofi, Menarini, AbbVie, Janssen, Oncopeptides, and GSK, and participated in advisory boards for AbbVie, Janssen, Regeneron, and Sanofi. The other authors declare no conflicts of interest., (© 2025 The Author(s). eJHaem published by British Society for Haematology and John Wiley & Sons Ltd.)

The intestinal epithelium has a remarkably high turnover in homeostasis. It remains unresolved how this is orchestrated at the cellular level and how the behavior of stem and progenitor cells ensures tissue maintenance. To address this, we combined quantitative fate mapping in three complementary mouse models with mathematical modeling and single-cell RNA sequencing. Our integrated approach generated a spatially and temporally defined model of crypt maintenance based on two cycling populations: stem cells at the crypt-bottom and transit-amplifying (TA) cells above them. Subsequently, we validated the predictions from the mathematical model, demonstrating that fate decisions between the secretory and absorptive lineages are made within the stem cell compartment, whereas TA cell divisions contribute specifically to the absorptive lineage. These quantitative insights provide further direct evidence for crypt-bottom stem cells as the dominant driver of the intestinal epithelium replenishment., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2024 The Author(s). Published by Elsevier Inc. All rights reserved.)

Objectives: This study aimed to investigate patients' use of electronic Patient-Reported Outcome Measures (ePROMs) and understand the demographic and clinical factors that may be correlated with patient responses to the BREAST-Q at the preoperative stage of breast cancer. The BREAST-Q is a PROM in questionnaire format, developed and validated to assess satisfaction and quality of life for breast surgery patients.The hypothesis tested is that considering disparities in geography, age and education among responders is essential for capturing a diverse patient population in future Patent-Reported Outcome Measures initiatives, examining how these characteristics are associated with Patent-Reported Outcome Measures utilisation and outcomes., Design: Quantitative descriptive study., Setting: Electronic Patient-Reported Outcome Measures were collected between 6 September 2021 and 5 September 2022 from patients recruited from an outpatient clinic at a Plastic- and Breast Surgery Department at a University Hospital in Denmark., Participants: Participants include a total of 629 Danish-speaking women diagnosed with breast cancer and scheduled for breast cancer surgery, with a final participation rate of 468., Intervention: Preoperative ePROMs and demographic data were collected between September 2020 and 2021 through patients' secure national digital post-box., Main Outcome Measures: Demographic variables of both responders and non-responders were assessed using t-tests, Mann-Whitney U tests and χ 2 tests. Linear regression models were employed to determine the demographic variables associated with BREAST-Q subscale scores., Results: The response rate for ePROMs was 72.5% with a median age of responders at 62 years. Older patients reported lower breast satisfaction (unadjusted coefficient b u =-0.26 (95% CI -0.44; -0.07), p=0.006) but better physical well-being (adjusted coefficient b a =0.23 (0.08; 0.37), p<0.001). Lower educational achievement was correlated with reduced breast satisfaction and psychosocial and sexual well-being; for example, patients with a master's/doctoral level education scored 14.29 points higher in psychosocial well-being (95% CI 6.50; 22.07, p<0.001) compared with those with lower secondary education. Cohabiting patients reported psychosocial well-being scores approximately four points higher than those living alone (b a =3.91 (0.06; 7.75), p=0.046). Body mass index (BMI) was negatively associated with sexual well-being, with a 0.75-point decline per additional BMI point (b a =-0.75, (-1.12; -0.37), p<0.001)., Conclusions: The present study demonstrates a positive attitude towards completing BREAST-Q as ePROMs among women diagnosed with breast cancer in the investigated region in Denmark. However, completion rates for ePROMs varied by demographic factors such as age, marital status and access to healthcare. Younger, more educated, married patients with lower BMI who lived near major cities were more likely to report better pretreatment outcomes., Competing Interests: Competing interests: None declared., (© Author(s) (or their employer(s)) 2025. Re-use permitted under CC BY-NC. No commercial re-use. See rights and permissions. Published by BMJ Group.)

Purpose: This study aimed to investigate patients' experiences with electronic Patient-Reported Outcome Measures (ePROMs) during follow-up consultations with registered nurses and surgeons in breast cancer care., Methods: A qualitative approach was employed, using focused ethnography with participant observations during patient consultations at a Plastic and Breast Surgery outpatient clinic, followed by individual interviews with the patients. Data were analysed using reflexive thematic analysis by Braun and Clarke. The discussion was informed by theory on person-centred practice., Results: A total of 38 participants were included. ePROMs strengthened person-centred practice by nudging patients to reflect on their health, voice their concerns and engage in active dialogue during consultations, particularly regarding body image issues. The relevance and impact of ePROMs were highly dependent on the timing of the patient's treatment trajectory. Patients found that ePROMs were more meaningful and contextually appropriate when completed during late follow-up. This could be because patients faced challenges with ePROMs, including not understanding their purpose, the need for assistance from relatives and instances when ePROMs were not integrated into consultations., Conclusion: ePROMs can empower patients by facilitating meaningful discussions about body image and other concerns during postoperative follow-up consultations. However, their effectiveness relies on clear communication and proper integration into clinical practice. The results add to existing litterature by highlighting the dual impact of ePROMs, as they can enhance patient-centred care but also present challenges when their purpose is unclear. The implications for future practice include the need to develop strategies that engage diverse patient groups in the ePROM process, supporting equal access to follow-up care and addressing health disparities., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Background and Aims: Epidemiological studies have shown that subnormal levels of vitamin D (25[OH]D) are associated with a more aggravated clinical course of ulcerative colitis [UC]. Despite an increased focus on the therapeutic importance of vitamin D and vitamin D receptor [VDR] signalling, the mechanisms underlying the effects of the vitamin D-VDR axis on UC remain elusive. Therefore, we aimed to investigate whether exposure to active vitamin D (1,25[OH]2D3/VDR) signalling in human organoids could influence the maintenance of the colonic epithelium., Methods: Intestinal VDR expression was studied by immunohistochemistry, RNA expression arrays, and single-cell RNA sequencing of colonic biopsy specimens obtained from patients with UC and healthy individuals. To characterise the functional and transcriptional effects of 1,25[OH]2D3, we used patient-derived colonic organoids. The dependency of VDR was assessed by knocking out the receptor with CRISPR/Cas9., Results: Our results suggest that 1,25[OH]2D3/VDR stimulation supports differentiation of the colonic epithelium and that impaired 1,25[OH]2D3/VDR signalling thereby may compromise the structure of the intestinal epithelial barrier, leading to flares of UC. Furthermore, a transcriptional response to VDR activity was observed primarily in fully differentiated cells at the top of the colonic crypt, and this response was reduced during flares of UC., Conclusions: We identified an important role of vitamin D signalling in supporting differentiated cell states in the human colonic epithelium, and thereby maintenance of the intestinal barrier integrity. This makes the vitamin D-VDR signalling axis an interesting target for therapeutic efforts to achieve and maintain remission in patients with UC., (© The Author(s) 2024. Published by Oxford University Press on behalf of European Crohn’s and Colitis Organisation.)

Introduction: The Patient Activity Treatment Outcome Scale (PATOS) is a novel patient-reported outcome measure (PROM). This study explored patients' and orthopaedic surgeons' experiences with PATOS as part of a PROM battery. It aimed to investigate its influence on patient involvement and healthcare decisions about knee or hip osteoarthritis patients scheduled for knee/hip arthroplasty., Methods: A qualitative research design was adopted using semi-structured interviews with 19 patients and nine orthopaedic surgeons at a Danish orthopaedic surgery department. Thematic analysis was used to develop key themes from interview transcripts., Results: This study adds nuanced insights related to the use of PROMs in routine orthopaedic clinical practice. Home-based completion of PROMs prompted patients to reflect on their situations, priorities and goals regarding potential hip or knee arthroplasty, leading to better-prepared discussions with surgeons. The integration of PATOS with other PROMs made it challenging to discuss PATOS exclusively. Therefore, the results relate to the full PROM battery. The experiences of patients and surgeons were organised into four themes: 1) Patient involvement is key, 2) Questionnaire load, 3) Meaningful home-based completion and 4) PROMs were not used in the decision-making process., Conclusions: The questionnaire load and limited integration into healthcare decision processes raised concerns about the application of the PROM battery. The results emphasise the importance of refining the application of PROMs in orthopaedic practice from the perspectives of patients and surgeons alike., Funding: This work was supported by Innovation Fund Denmark [0172-01258B] TRIAL REGISTRATION. Not relevant., (Published under Open Access CC-BY-NC-BD 4.0. https://creativecommons.org/licenses/by-nc-nd/4.0/.)

Background: Eosinophilic oesophagitis (EoE) is a chronic immune-mediated disease. In Denmark, the budesonide orodispersible tablet (BOT) is recommended as a second-line treatment for proton pump inhibitor-refractory EoE patients., Aims: To evaluate the effectiveness of treatment with BOT in adult EoE patients in a population-based setting in Denmark., Methods: This was a retrospective, registry-based, DanEoE cohort study of all 76 adult EoE patients treated with BOT and diagnosed between 2007 and 2021 in the North Denmark Region. After medical record revision, the EoE diagnosis was defined according to the AGREE consensus. Symptomatic response was based on the information found in the patients' medical reports and histologic remission was defined as <15 eosinophils per high-power field (eos/hpf)., Results: Histologic remission was achieved in 89% of the patients treated with BOT who underwent histologic evaluation. Clinicohistologic remission was achieved in 71% of the patients who underwent both symptomatic and histologic evaluation. Despite histologic remission, 18% of patients still experienced symptoms. Non-responders were found in 7% of the patients. Complications were rare, with dilation of strictures performed in 7% and food bolus obstruction (FBO) occurring in 3%. Discontinuation of the treatment due to unacceptable side effects was observed in 11% of the treated patients., Conclusions: Treatment with BOT effectively induced histologic remission in most of the EoE patients. Despite achieving histologic remission, approximately 1/5 of the patients were still symptomatic. Complications were rare. In non-responders and those with unacceptable side effects, alternative treatment options such as biologic agents might be needed.

The intestinal epithelium ensures uptake of vital nutrients and acts as a barrier between luminal contents and the underlying immune system. In inflammatory bowel diseases, such as ulcerative colitis (UC), this barrier is compromised, and patients experience debilitating symptoms. Here, we perform single-cell RNA profiling of epithelial cells and outline patterns of cell fate decisions in healthy individuals and UC patients. We demonstrate that patterns of hierarchical behavior are altered in UC patients and identify unique cellular states associated with Janus kinase/signal transducer and activator of transcription (JAK/STAT) activation in ulcerated and non-ulcerated areas of the colonic epithelium. These transcriptional changes could be recapitulated in human colonic organoids, wherein cytokine-mediated activation of JAK/STAT led to the emergence of cell populations with augmented regenerative properties. Altogether, our findings indicate that intricate relationships between epithelial and cytokine signaling regulate cell fate during epithelial tissue regeneration in humans and have important implications for the understanding of UC biology., Competing Interests: Declaration of interests T.S. is an inventor on several patents related to organoids., (Copyright © 2024 The Author(s). Published by Elsevier Inc. All rights reserved.)

Neurodegenerative diseases constitute a global health issue and a major economic burden. They significantly impair both cognitive and motor functions, and their prevalence is expected to rise due to ageing societies and continuous population growth. Conventional therapies provide symptomatic relief, nevertheless, disease-modifying treatments that reduce or halt neuron death and malfunction are still largely unavailable. Amongst the common hallmarks of neurodegenerative diseases are protein aggregation, oxidative stress, neuroinflammation and mitochondrial dysfunction. Transcription factor nuclear factor-erythroid 2-related factor 2 (NRF2) constitutes a central regulator of cellular defense mechanisms, including the regulation of antioxidant, anti-inflammatory and mitochondrial pathways, making it a highly attractive therapeutic target for disease modification in neurodegenerative disorders. Here, we describe the role of NRF2 in the common hallmarks of neurodegeneration, review the current pharmacological interventions and their challenges in activating the NRF2 pathway, and present alternative therapeutic approaches for disease modification., Competing Interests: CM, LR-P, JE, and SH are employees and HK and SK are scientific cofounders of NEUmiRNA Therapeutics, a biopharmaceutical company developing RNA-based therapeutics for neurological disorders., (Copyright © 2024 Mayer, Riera-Ponsati, Kauppinen, Klitgaard, Erler and Hansen.)

This study investigated the efficacy of a 10-session nature-based therapeutic intervention for people with post-concussion symptoms. The intervention involved physical and vestibular exercises, sensory training, relaxation, and psychoeducation, all of which were integrated with the natural environment in a forest therapy garden. This study was designed with a passive control period followed by the intervention ( n = 30). The Mental Fatigue Scale (MFS) was the primary outcome measure. The secondary outcome measures were the Warwick-Edinburg Mental Wellbeing Scale and the short version of the Quality of Life after Brain Injury. A Likert scale was used to examine the mental strain of the sessions themselves. The MFS (primary outcome) exhibited a significant decrease with a medium-sized effect from before to after the intervention. The secondary outcomes exhibited significant increases from the beginning to the end of the intervention. All outcomes were sustained at follow-up ten weeks later. No significant difference was found from the control period. This study indicates that the described nature-based intervention is a feasible treatment for reducing prolonged post-concussion symptoms. However, it should be studied more in-depth to understand the impact of the natural environment and to validate the results on a larger representative population.

Background: Antipsychotics are commonly administered to nursing home residents with dementia, despite the associated risk of severe adverse events., Objective: This study aimed to explore healthcare professionals' experiences in caring for nursing home residents with dementia, with a focus on rationales behind the use of antipsychotics., Method: Twelve semi-structured interviews with healthcare professionals' from Danish nursing homes were conducted and analyzed using the method Systematic Text Condensation., Results: Nonpharmacological interventions were reported as the primary approach to care and the first-choice treatment for behavioral and psychological symptoms of dementia (BPSD). Use of antipsychotics was considered to serve as a last resort, reserved for residents with severe symptoms. However, most informants preferred a more limited use. The study identified four main barriers to reduce the use of antipsychotics: "Scarcity of resources", "Perceiving antipsychotic use to provide relieve", "Reluctance towards deprescribing" and "Limited access to medical counseling", and three potential enablers: " Updating knowledge and nonpharmacological competencies ", " Management support and clear procedures " and " Regularity in interdisciplinary collaboration "., Conclusion: The treatment and care were reported as primarily following guidelines in BPSD. Several barriers were perceived to challenge the healthcare professionals' preference of limited use of antipsychotics. To further reduce the use, this study highlights the importance of understanding the adverse effects caused by limited resources, enhancing employee knowledge and competencies and ensuring regular interprofessional collaboration for assessing and reassessing the need to use antipsychotics., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. Charlotte Vermehren received a grant by the Danish Health Authority (3165–0018) to fund the DECADE-project. The Danish Health Authority was not involved in study decisions. No other conflict of interest or funding relevant to the current study were declared., (© 2024 The Authors.)

This is a case report of two men aged 39 and 43 years with dissection of the coeliac trunk involving the splenic arteries causing splenic infarction. One case was associated with an increase in abdominal pressure during defaecation and the other occurred during treatment with methylphenidate. Based on the published 43 cases, risk factors include male sex, increased intraabdominal pressure or increased vascular pressure. Methylphenidate most likely increased the blood pressure, and dissections of other arteries have been described during treatment with this and the similar drug amphetamine., (Published under Open Access CC-BY-NC-BD 4.0. https://creativecommons.org/licenses/by-nc-nd/4.0/.)

Objectives: This review aims to analyse the published data on preclinical and human experimental and clinical adenosine modulation for pain management. We summarise the translatability of the adenosine pathway for further drug development and aim to reveal subgroups of pain patients that could benefit from targeting the pathway., Content: Chronic pain patients suffer from inadequate treatment options and drug development is generally impaired by the low translatability of preclinical pain models. Therefore, validating the predictability of drug targets is of high importance. Modulation of the endogenous neurotransmitter adenosine gained significant traction in the early 2000s but the drug development efforts were later abandoned. With the emergence of new drug modalities, there is a renewed interest in adenosine modulation in pain management. In both preclinical, human experimental and clinical research, enhancing adenosine signalling through the adenosine receptors, has shown therapeutic promise. A special focus has been on the A 1 and A 3 receptors both of which have shown great promise and predictive validity in neuropathic pain conditions., Summary: Adenosine modulation shows predictive validity across preclinical, human experimental and clinical investigations. The most compelling evidence is in the field of neuropathic pain, where adenosine has been found to alleviate hyperexcitability and has the potential to be disease-modifying., Outlook: Adenosine modulation show therapeutic potential in neuropathic pain if selective and safe drugs can be developed. New drug modalities such as RNA therapeutics and cell therapies may provide new options., (© 2023 the author(s), published by De Gruyter, Berlin/Boston.)

Drug discovery in epilepsy began with the finding of potassium bromide by Sir Charles Locock in 1857. The following century witnessed the introduction of phenotypic screening tests for discovering antiseizure medications (ASMs). Despite the high success rate of developing ASMs, they have so far failed in eliminating drug resistance and in delivering disease-modifying treatments. This emphasizes the need for new drug discovery strategies in epilepsy. RNA-based drugs have recently shown promise as a new modality with the potential of providing disease modification and counteracting drug resistance in epilepsy. RNA therapeutics can be directed either toward noncoding RNAs, such as microRNAs, long noncoding RNAs (ncRNAs), and circular RNAs, or toward messenger RNAs. The former show promise in sporadic, nongenetic epilepsies, as interference with ncRNAs allows for modulation of entire disease pathways, whereas the latter seem more promising in monogenic childhood epilepsies. Here, we describe therapeutic strategies for modulating disease-associated RNA molecules and highlight the potential of RNA therapeutics for the treatment of different patient populations such as sporadic, drug-resistant epilepsy, and childhood monogenic epilepsies., (© 2023 International League Against Epilepsy.)

The intestinal epithelium is constantly exposed to microbes residing in the lumen. Traditionally, the response to microbial interactions has been studied in cell lines derived from cancerous tissues, e.g. Caco-2. It is, however, unclear how the responses in these cancer cell lines reflect the responses of a normal epithelium and whether there might be microbial strain-specific effects. To address these questions, we derived organoids from the small intestine from a cohort of healthy individuals. Culturing intestinal epithelium on a flat laminin matrix induced their differentiation, facilitating analysis of microbial responses via the apical membrane normally exposed to the luminal content. Here, it was evident that the healthy epithelium across multiple individuals ( n  = 9) demonstrates robust acute both common and strain-specific responses to a range of probiotic bacterial strains (BB-12 Ⓡ , LGG Ⓡ , DSM33361, and Bif195). Importantly, parallel experiments using the Caco-2 cell line provide no acute response. Collectively, we demonstrate that primary epithelial cells maintained as organoids represent a valuable resource for assessing interactions between the epithelium and luminal microbes across individuals, and that these models are likely to contribute to a better understanding of host microbe interactions.

Introduction: Behavioural and psychological symptoms of dementia (BPSD) should only rarely and briefly be treated with antipsychotics. Despite recommendations to the contrary, the use of antipsychotics in nursing home residents with dementia is widespread and followed by serious adverse effects. Intervention studies on methods to reduce the use of antipsychotics in persons with dementia are few and needed. The aim of this protocol is to describe the rationale and content of the intervention DEprescribing and Care to reduce Antipsychotics in DEmentia (DECADE)-a hybrid effectiveness-implementation pilot study., Materials and Methods: This is a protocol of a prospective hybrid effectiveness-implementation pilot study. The primary aim of DECADE is to reduce the use of antipsychotic drugs by 50% in 50% of nursing home residents with dementia while maintaining or improving BPSD. The intervention is implemented in six nursing homes including approximately 190 residents with dementia and consists of Academic Detailing, medication review, education of nursing home staff, and care plans. The evaluation of feasibility and potential effectiveness is an overall assessment of all clinical and process outcomes. Logistic regression analyses will be used to investigate factors characterizing situations with prescription of antipsychotics. BPSD is analysed with a before- and after design using self-controlled case series methods and the use of antipsychotics is analysed as interrupted time series., Discussion: This protocol describes a study that will provide an indication of DECADE effectiveness and a model for upscaling and further evaluation in a controlled design., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2023 Benthien et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Human IVF embryos that are not used for fresh transfer are cryopreserved by vitrification for later embryo transfers. This study evaluates pre-vitrification and post-warming embryo characteristics that are suitable to predict the chance of clinical pregnancy in single vitrified blastocyst transfer (SVBT) cycles. In a multicenter observational trial (IMBOS trial), embryos were cultured in a time-lapse system before and after vitrification. Associations between clinical pregnancy, morphokinetic parameters, blastocyst collapse, KIDScore D5, pre-vitrification and post-warming Gardner scores, post-warming blastocyst size and re-expansion rates before SVBT were analyzed in 182 SVBTs which resulted in 89 clinical pregnancies. No association was found between clinical pregnancy after SVBT and the number of collapses or the maximal collapse size before vitrification. The multifactorial analysis of pre-vitrification Gardner scores showed a significant association with clinical pregnancy for trophectoderm grading but not for expansion/hatching status and inner cell mass grading. A significant association with clinical pregnancy was found for the time to reach a blastocyst after pronuclear fading (tB-tPNf), KIDScore D5 and post-warming size but not the rate of expansion or maximal expansion size. The selection of blastocysts for SVBT could benefit from using pre-vitrification parameters like tB-tPNf, trophectoderm grading and post-warming blastocyst size.

A major barrier to applying deep segmentation models in the medical domain is their typical data-hungry nature, requiring experts to collect and label large amounts of data for training. As a reaction, prototypical few-shot segmentation (FSS) models have recently gained traction as data-efficient alternatives. Nevertheless, despite the recent progress of these models, they still have some essential shortcomings that must be addressed. In this work, we focus on three of these shortcomings: (i) the lack of uncertainty estimation, (ii) the lack of a guiding mechanism to help locate edges and encourage spatial consistency in the segmentation maps, and (iii) the models' inability to do one-step multi-class segmentation. Without modifying or requiring a specific backbone architecture, we propose a modified prototype extraction module that facilitates the computation of uncertainty maps in prototypical FSS models, and show that the resulting maps are useful indicators of the model uncertainty. To improve the segmentation around boundaries and to encourage spatial consistency, we propose a novel feature refinement module that leverages structural information in the input space to help guide the segmentation in the feature space. Furthermore, we demonstrate how uncertainty maps can be used to automatically guide this feature refinement. Finally, to avoid ambiguous voxel predictions that occur when images are segmented class-by-class, we propose a procedure to perform one-step multi-class FSS. The efficiency of our proposed methodology is evaluated on two representative datasets for abdominal organ segmentation (CHAOS dataset and BTCV dataset) and one dataset for cardiac segmentation (MS-CMRSeg dataset). The results show that our proposed methodology significantly (one-sided Wilcoxon signed rank test, p<0.05) improves the baseline, increasing the overall dice score with +5.2, +5.1, and +2.8 percentage points for the CHAOS dataset, the BTCV dataset, and the MS-CMRSeg dataset, respectively., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Author(s). Published by Elsevier B.V. All rights reserved.)

Background and Aim: The DanEoE is a previously described population- and register-based cohort of 236 adult patients with eosinophilic esophagitis (EoE) in a well-defined Danish region with a population of 580 000 and free medical treatment. The aim of the study was to compare the phenotype and treatment response between EoE patients with complications to patients without complications at diagnosis., Methods: A retrospective cross-sectional study of the DanEoE cohort's 236 adult EoE patients diagnosed between 2007 and 2017 in the North Denmark Region. Patients were divided into a group who had had complications (dilated or food bolus obstruction [FBO]) before or at the diagnosis, and a group without., Results: At the diagnostic endoscopy, 61% had never had a complication, and 39% had either had FBO ( n  = 77) or been dilated ( n  = 15). The complicated group had the same mean age at symptom debut (37 [SD = 16] vs 37 [SD = 17] years, P  = 1.0), but were diagnosed significantly later with a resulting longer diagnostic delay (13 [SD = 13] vs 7.9 [SD = 11] years, P  = 0.01). Almost half of all patients were never treated to symptomatic remission (uncomplicated 40%, complicated 49%). The histological remission was not secured in the majority (uncomplicated 68%, complicated 70%). Despite this, <15% of patients with previous FBO experienced this after the diagnosis., Conclusion: In the population-based DanEoE cohort, results indicated that the complicated EoE phenotype was a patient with a 5-year longer diagnostic delay. In the current study, the complication status did not predict the treatment response., (© 2023 The Authors. JGH Open published by Journal of Gastroenterology and Hepatology Foundation and John Wiley & Sons Australia, Ltd.)

During intestinal organogenesis, equipotent epithelial progenitors mature into phenotypically distinct stem cells that are responsible for lifelong maintenance of the tissue. While the morphological changes associated with the transition are well characterized, the molecular mechanisms underpinning the maturation process are not fully understood. Here, we leverage intestinal organoid cultures to profile transcriptional, chromatin accessibility, DNA methylation, and three-dimensional (3D) chromatin conformation landscapes in fetal and adult epithelial cells. We observed prominent differences in gene expression and enhancer activity, which are accompanied by local changes in 3D organization, DNA accessibility, and methylation between the two cellular states. Using integrative analyses, we identified sustained Yes-Associated Protein (YAP) transcriptional activity as a major gatekeeper of the immature fetal state. We found the YAP-associated transcriptional network to be regulated at various levels of chromatin organization and likely to be coordinated by changes in extracellular matrix composition. Together, our work highlights the value of unbiased profiling of regulatory landscapes for the identification of key mechanisms underlying tissue maturation.

Generation of functionally mature organs requires exquisite control of transcriptional programs governing cell state transitions during development. Despite advances in understanding the behavior of adult intestinal stem cells and their progeny, the transcriptional regulators that control the emergence of the mature intestinal phenotype remain largely unknown. Using mouse fetal and adult small intestinal organoids, we uncover transcriptional differences between the fetal and adult state and identify rare adult-like cells present in fetal organoids. This suggests that fetal organoids have an inherent potential to mature, which is locked by a regulatory program. By implementing a CRISPR-Cas9 screen targeting transcriptional regulators expressed in fetal organoids, we establish Smarca4 and Smarcc1 as important factors safeguarding the immature progenitor state. Our approach demonstrates the utility of organoid models in the identification of factors regulating cell fate and state transitions during tissue maturation and reveals that SMARCA4 and SMARCC1 prevent precocious differentiation during intestinal development.

Glucagon-like peptide 1 (GLP-1) receptor agonists can decrease alcohol intake by central mechanisms that are still poorly understood. The lateral septum (LS) and the ventral/caudal part of the hippocampus are enriched in GLP-1 receptors, and activity in these regions was shown to modulate reward-related behaviors. Using microinfusions of the GLP-1 receptor agonist exendin-4 in mice trained to self-administer oral alcohol in an operant assay, we tested whether pharmacological stimulation of GLP-1 receptors in hippocampus and LS decrease alcohol self-administration. We report that infusion of exendin-4 in the ventral hippocampus or LS was sufficient to reduce alcohol self-administration with as large effect sizes as we previously reported with systemic exendin-4 administration. Infusion of exendin-4 into the nucleus accumbens also reduced alcohol self-administration, as anticipated based on earlier reports, while infusion of exendin-4 into the caudate-putamen (dorsal striatum) had little effect, consistent with lack of GLP-1 receptor expression in this region. The distribution of exendin-4 after infusion into the LS or caudate putamen was visualized using a fluorescently labeled ligand. These findings add to our understanding of the circuit-level mechanisms underlying the ability of GLP-1 receptor agonists to reduce alcohol self-administration. (PsycInfo Database Record (c) 2023 APA, all rights reserved).

Background: Few studies are available regarding occupational therapist students' experiences relating to their professional identity during their education., Aim: The aim was to gain knowledge about the process that occupational therapist students undergo in the shaping of their professional identity., Method: Semi-structured interviews were conducted with nine participants divided into two phases: a phenomenological phase, followed up by a hermeneutical phase. The data was analysed with the use of thematic analysis., Results: Three themes emerged: study environment, responsibility, and choice of internships. As a part of the first theme study environments, the relations among the students were important for the shaping of a professional identity., Conclusion: The occupational therapy students undergo an increasing sense of responsibility throughout the education programme. Particularly, the clinical practice was found to have a positive impact on the shaping of professional identity., Important Findings: The findings in the study can contribute to clarifying students' perspectives on the shaping of their professional identity. Based on these findings the occupational therapist education programmes could integrate these elements as a part of their curriculum.

Drug concentrations in peripheral blood are often used to evaluate whether death was caused by drug intoxication. In some cases, peripheral blood is not available, and analytical results of alternative matrices should instead be used in the toxicological evaluation. However, reference concentrations of alternative matrices are few, which makes interpretation of results a challenge. In this study, concentrations of selected benzodiazepines, opioids, illicit drugs, and other commonly used drugs in postmortem femoral blood, cardiac blood, brain tissue, and muscle tissue are presented. Alternative matrix-to-femoral blood drug concentration ratios and correlations of blood and alternative matrix drug concentrations were calculated to examine which of the investigated alternative matrices were most suited to use for toxicological evaluation in cases where peripheral blood is not available. The results showed that concentrations in cardiac blood, brain tissue, and muscle tissue could be useful in the postmortem evaluation of most of the 19 selected analytes. In most cases, analytes were detected in all the alternative matrices. The median concentration ratios for the selected analytes in brain tissue, cardiac blood, and muscle tissue relative to femoral blood ranged from 0.57 to 3.42, 0.59 to 1.87, and 0.67 to 7.04, respectively. Overall, cardiac blood provided the concentrations most comparable with femoral blood concentrations, indicating that cardiac blood can be useful in cases where femoral blood is not available. However, the measured concentrations should be interpreted with caution., (© 2023 The Authors. Drug Testing and Analysis published by John Wiley & Sons Ltd.)

Background: Symptoms of stress are widespread in the working population and associated with long-term sickness absence and poor work functioning. Occupational therapy (OT)-based interventions are effective in improving return-to-work (RTW)-rates in stress-afflicted long-term sickness benefit beneficiaries (SBBs). No Danish standard on OT-RTW for stress-afflicted SSBs exists., Aims: To describe and evaluate the feasibility of a seven-week OT-RTW intervention for stress-afflicted SBBs., Materials and Methods: Daily Life Coping (DLC) utilizes peer-learning and salutogenetic perspectives to address occupational imbalances experienced by long-term SBBs. Stress-afflicted SBBs ( n  = 54) in the municipality of Hilleroed were enrolled. Self-efficacy and well-being were measured before and after intervention using the general self-efficacy (GSE) scale and WHO-5 Well-Being Index (WHO-5). RTW-rates were measured at one-year follow-up. Participant feedback was collected using standardized evaluation forms., Results: Following the intervention, WHO-5 improved by 13.7 ± 16.5 points and GSE improved by 4.3 ± 4.8 points. Participant feedback was positive. At one-year follow-up, employment status data were available for 51 SSBs of these 58.8% achieved employment. High post-test GSE and WHO-5 scores were associated with employment at follow-up., Conclusion: DLC led to significant improvements in self-efficacy and well-being. Preliminary results indicate benefits on RTW-rates., Significance: This study provides evidence of the feasibility of DLC in a municipal setting.

Alternative specimens collected during autopsies can be valuable in postmortem toxicology in cases where peripheral blood is not available. The applicability of brain tissue as an alternative matrix for drug screening by ultra-high-performance liquid chromatography-quadrupole time-of-flight mass spectrometry was investigated in this study. Results of the 50 most frequently detected drugs and metabolites of toxicological interest in blood and brain tissue samples from 1,719 autopsy cases were compared. Examination of the results in paired blood and brain tissue samples revealed that the two matrices were in general comparable, as the majority of the 50 analytes were observed in a high number of the examined cases in both blood and brain tissue. This demonstrates the potential of brain tissue as an alternative matrix for drug screening in postmortem toxicology or as a secondary matrix for confirmation., (© The Author(s) 2022. Published by Oxford University Press.)

The expeditious progress of Mesenchymal Stromal Cells (MSC) for therapeutic intervention calls for means to compare differences in potency of cell products. The differences may be attributed to innumerable sources including tissue origin, production methods, or even between batches. While the immunomodulatory potential of MSC is recognized and well-documented by an expansive body of evidence, the methodologies and findings vary markedly. In this study, we utilized flowcytometric analysis of lymphocyte proliferation based on cryopreserved peripheral blood mononuclear cells for quantification of the inhibitory effect of MSC. Technical aspects of fluorescent staining and cryopreservation of peripheral blood mononuclear cells were evaluated to obtain optimal results and increase feasibility. A range of common specific and unspecific mitogens was titrated to identify the conditions, in which the effects of Adipose tissue-derived Stromal Cells (ASC; a type of MSC) were most pronounced. Specific stimulation by antibody-mediated activation of CD3 and CD28 via TransAct and Dynabeads lead to substantial proliferation of lymphocytes, which was inhibited by ASC. These results were closely mirrored when applying unspecific stimulation in form of phytohemagglutinin (PHA), but not concanavalin A or pokeweed mitogen. The mixed lymphocyte reaction is a common assay which exploits alloreactivity between donors. While arguably more physiologic, the output of the assay often varies substantially, and the extent of proliferation is limited since the frequency of alloreactive cells is low, as opposed to the mitogens. To heighten the proliferative response and robustness, combinations of 2-5 donors were tested. Maximum proliferation was observed when combining 4 or more donors, which was efficiently suppressed by ASC. Several desirable and unfavorable traits can be attributed to the tested stimuli in the form of keywords. The importance of these traits should be scored on a laboratory-level to identify the ideal mitogen. In our case the ranking listed PHA as the most suited candidate. Developing robust assays is no trivial feat. By disclosing the full methodological framework in the present study, we hope to aid others in establishing functional metrics on the road to potency assays., Competing Interests: Authors AE and JK are inventors of the patent “Stem cell therapy based on adipose-derived stem cells” Publication WO 2017-068140 filed by Rigshospitalet, Denmark. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Hansen, Højgaard, Kastrup, Ekblond, Follin and Juhl.)

Introduction: The use of patient-reported outcome measures (PROMs) in clinical practice has the potential to promote person-centred care and improve patients' health-related quality of life. We aimed to develop an intervention centred around electronic PROMs (ePROMs) for systematic follow-up in patients diagnosed with breast cancer and to evaluate its feasibility., Methods and Analysis: We developed a nurse-oriented and surgeon-oriented intervention in PROMs, including (1) an education programme for nurses and surgeons; (2) administration of BREAST-Q as proactive ePROMs during follow-up in patients diagnosed with breast cancer and (3) feedback to nurses and surgeons on PROM scores and a guidance manual for healthcare practitioners. Subsequently, we designed a non-controlled feasibility evaluation on the outcomes acceptability, demand, implementation, practicality and integration. The feasibility evaluation includes qualitative ethnographic studies exploring the user perspectives of patients, nurses and surgeons and quantitative studies to explore the characteristics of the patient population regarding demographic background, response rates and response patterns. The feasibility study was initiated in September 2021, will continue until 2024 and will include approximately 900 patients. EPROMs are collected at the following assessment time points: baseline (after diagnosis, before surgery), 1-year follow-up and 3-year endpoint., Ethics and Dissemination: The study will be conducted according to the General Data Protection Regulation and the fifth version of the Helsinki Declaration. The National Committee on Health Research Ethics approved the study according to the law of the Committee § 1, part 4. All data will be anonymised before its publication. The results of the feasibility study will be published in peer-reviewed, international journals., Competing Interests: Competing interests: None declared., (© Author(s) (or their employer(s)) 2022. Re-use permitted under CC BY-NC. No commercial re-use. See rights and permissions. Published by BMJ.)

Aims: This study explores how women diagnosed with breast cancer may be supported by physicians and nurses during physical and existential changes related to oncoplastic breast surgery in Denmark. The following research questions were addressed: (a) how do women experience oncoplastic breast surgery, and (b) how does cancer treatment affect their body image?, Design: A descriptive qualitative study design with a six-step thematic analysis influenced by Braun and Clarke was applied in this study. This paper has been prepared in accordance with the consolidated criteria for reporting qualitative research., Methods: Fourteen in-depth interviews with seven women diagnosed with breast cancer were conducted from August 2018 to March 2019. In this qualitative study, data analysis was performed concurrent with data construction, recognizing that the process of analysis and making sense of data should start during the interviews. We explicitly frame the discussion of the findings in a theory of embodiment influenced by Merleau-Ponty, consistent with the construct of exploring human experiences to generate meaningful knowledge for applied practice., Results: Two overall themes with related subthemes were identified: (1) 'Treatment is required for life-threatening cancer', and (2) 'Striving for a new normal body'. Across both themes, women's experiences reflected a 'time pendulum' as they contemplated their past identity, their current rationale and their transition to a future beyond breast cancer with a changed body., Conclusion: Participants reflected on their past, present and future when facing an altered body image caused by their breast cancer diagnosis and oncoplastic breast surgery. The participants in the study expressed broad levels of satisfaction with the results of the oncoplastic breast surgery. The reconstructed breast helped them to live normally again, in particular maintaining interpersonal relationships. Breast reconstruction supported participants' embodiment experiences and redefinition of their 'new normal'., Impact: This study showed the dynamic changes in self-definition from receiving a breast cancer diagnosis and cancer treatment to oncoplastic breast surgery. The main finding of self-redefinition was from the perspective of breast cancer women who were in a period of transition between post-diagnosis and consultation for oncoplastic breast surgery. The findings indicate that advanced nurse specialists in the field of oncoplastic breast surgery can enhance psychosocial wellbeing and support women pre- and post-operatively by focusing on patient experiences of self-image and embodiment., (© 2022 The Authors. Journal of Advanced Nursing published by John Wiley & Sons Ltd.)

Precision (or imprecision) is one of the central performance parameters of all analytical methods. It is often evaluated during method validation in spiked samples, with a relatively low number of measurements typically during a short period. Validation results are well documented in the literature; however, evaluation of imprecision in authentic cases compared with long-term imprecision from quality control samples has not often been reported on. The aim of this study was to investigate imprecision estimated from duplicate measurements of ante- and postmortem blood samples and long-term imprecision estimates from quality control samples and to compare variations between them. Data were analyzed by using robust statistics, where results for the 29 analytes most frequently quantified by liquid chromatography-tandem mass spectrometry in ante- and postmortem blood samples were included. A total of 41,460 positive findings in authentic whole blood and 51,522 measurements from quality controls were investigated. Analysis was performed in duplicate on independent batches on two separate days. Overall, the imprecision estimated in quality control and the authentic samples were akin for most analytes. Ante- and postmortem blood samples showed similar imprecision for the majority of the analytes and were approximately the same level as long-term imprecision estimated from the quality control samples at low level, whereas relative imprecision of the quality control samples at high level were slightly lower than ante- and postmortem blood samples. The methods we evaluated showed satisfactory reproducibility and robustness for the investigated analytes., Competing Interests: Conflicts of Interest None., (Copyright © 2022 The Author(s). Published by Elsevier B.V. All rights reserved.)

Recent work has shown that label-efficient few-shot learning through self-supervision can achieve promising medical image segmentation results. However, few-shot segmentation models typically rely on prototype representations of the semantic classes, resulting in a loss of local information that can degrade performance. This is particularly problematic for the typically large and highly heterogeneous background class in medical image segmentation problems. Previous works have attempted to address this issue by learning additional prototypes for each class, but since the prototypes are based on a limited number of slices, we argue that this ad-hoc solution is insufficient to capture the background properties. Motivated by this, and the observation that the foreground class (e.g., one organ) is relatively homogeneous, we propose a novel anomaly detection-inspired approach to few-shot medical image segmentation in which we refrain from modeling the background explicitly. Instead, we rely solely on a single foreground prototype to compute anomaly scores for all query pixels. The segmentation is then performed by thresholding these anomaly scores using a learned threshold. Assisted by a novel self-supervision task that exploits the 3D structure of medical images through supervoxels, our proposed anomaly detection-inspired few-shot medical image segmentation model outperforms previous state-of-the-art approaches on two representative MRI datasets for the tasks of abdominal organ segmentation and cardiac segmentation., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 The Author(s). Published by Elsevier B.V. All rights reserved.)

RNA therapeutics comprise a diverse group of oligonucleotide-based drugs such as antisense oligonucleotides (ASOs), small interfering RNAs (siRNAs), and short hairpin RNAs (shRNAs) that can be designed to selectively interact with drug targets currently undruggable with small molecule-based drugs or monoclonal antibodies. Furthermore, RNA-based therapeutics have the potential to modulate entire disease pathways, and thereby represent a new modality with unprecedented potential for generating disease-modifying drugs for a wide variety of human diseases, including central nervous system (CNS) disorders. Here, we describe different strategies for delivering RNA drugs to the CNS and review recent advances in clinical development of ASO drugs and siRNA-based therapeutics for the treatment of neurological diseases and neuromuscular disorders. Abbreviations 2'-MOE: 2'- O -(2-methoxyethyl); 2'- O -Me: 2'- O -methyl; 2'-F: 2'-fluoro; AD: Alzheimer's disease; ALS: Amyotrophic lateral sclerosis; ALSFRS-R: Revised Amyotrophic Lateral Sclerosis Functional Rating Scale; ARC: Antibody siRNA Conjugate; AS: Angelman Syndrome; ASGRP: Asialoglycoprotein receptor; ASO: Antisense oligonucleotide; AxD: Alexander Disease; BBB: Blood brain barrier; Bp: Basepair; CNM: Centronuclear myopathies; CNS: Central Nervous System; CPP: Cell-penetrating Peptide; CSF: Cerebrospinal fluid; DMD: Duchenne muscular dystrophy; DNA: Deoxyribonucleic acid; FAP: Familial amyloid polyneuropathy; FALS: Familial amyotrophic lateral sclerosis; FDA: The United States Food and Drug Administration; GalNAc: N-acetylgalactosamine; GoF: Gain of function; hATTR: Hereditary transthyretin amyloidosis; HD: Huntington's disease; HRQOL: health-related quality of life; ICV: Intracerebroventricular; IT: Intrathecal; LNA: Locked nucleic acid; LoF: Loss of function; mRNA: Messenger RNA; MS: Multiple Sclerosis; MSA: Multiple System Atrophy; NBE: New Biological Entity; NCE: New Chemical Entity; NHP: Nonhuman primate; nt: Nucleotide; PD: Parkinson's disease; PNP: Polyneuropathy; PNS: Peripheral nervous system; PS: Phosphorothioate; RISC: RNA-Induced Silencing Complex; RNA: Ribonucleic acid; RNAi: RNA interference; s.c.: Subcutaneous; siRNA: Small interfering RNA; SMA: Spinal muscular atrophy; SMN: Survival motor neuron; TTR: Transthyretin.

Objectives: In the current study, we aimed to explore the experiences and attitudes among healthcare professionals as they transitioned from their familiar disciplines to respiratory medicine, intensive care or other departments during the first wave of the COVID-19 pandemic., Background: In preparation for the increasing number of patients suspected of having or who would be severely ill from COVID-19, a major reconstruction of the Danish Healthcare System was initiated. The capacity of the healthcare system to respond to the unprecedented situation was dependent on healthcare professionals' willingness and ability to engage in these new circumstances. For some, this may have resulted in uncertainty, anxiety and fear., Design: The study was a descriptive study using semi-structured focus group interviews. Healthcare professionals (n = 62) from seven departments were included, and 11 focus group interviews were conducted. The focus group interviews took place during June 2020. Analyses was conducted using thematic analysis. The current study was reported using the consolidated criteria for reporting Qualitative research (COREQ)., Results: Healthcare professionals experiences was described by five themes: 1) Voluntary involvement, 2) Changes within the organisation, 3) Risks, 4) Professional identity and 5) Personal investment. Common to all five themes was the feeling of being on a pendulum from a meaningful experience to an experience of mental overload, when situations and decisions no longer seemed to be worthwhile., Conclusions: Healthcare professionals experienced a pendulum between a meaningful experience and one of mental overload during the COVID-19 pandemic. The swinging was conditioned by the prevailing context and was unavoidable., Relevance to Clinical Practice: To balance the continuous pendulum swing, leaders must consider involvement, and to be supportive and appreciative in their leader style. This is consistent with a person-centred leadership that facilitates a well-adjusted work-life balance and may help prevent mental overload developing into burnout., (© 2021 John Wiley & Sons Ltd.)

Introduction: Health care systems worldwide are currently facing major challenges because of the coronavirus disease pandemic. When individuals experience coronavirus disease symptoms, they often have to decide whether to seek health care services and render themselves vulnerable to infection or stay home and monitor their condition. Coronavirus disease management strategies should aim to reduce transmission, promote disease control, and facilitate self-monitoring within the population. In this regard, mobile health technologies serve as supportive tools, and acquiring knowledge about user perspectives will facilitate the development and integration of coronavirus disease-related applications. Accordingly, this study aimed to examine user perspectives on applications that monitor coronavirus disease-related physical signs and identify discrepancies between user expectations and developer design perspectives within the Danish context., Materials and Methods: A qualitative research design was adopted. Semi-structured telephone interviews were conducted to examine user expectations during the first wave of the coronavirus disease pandemic in April 2020. The theoretical framework, which was inspired by the concept of health literacy, was developed using a six-step thematic analytic approach., Results: The analysis yielded two major themes that captured user experiences: (1) coronavirus disease-related applications may serve as digital tools that foster safety when physical signs are monitored and (2) the acceptability of coronavirus disease-related applications depends on the adoption of a personalised and user-friendly design., Competing Interests: Declaration of conflicting interests: The author(s) declared no potential conflicts of interest with respect to the research, authorship and/or publication of this article., (© The Author(s) 2021.)

Whole blood is most often the matrix of choice for postmortem analysis but it is not always available. In these cases, muscle tissue can be used as an alternative matrix. Therefore, an ultra-high-performance liquid chromatography-tandem mass spectrometry method for the quantification of 29 drugs and metabolites of toxicological interest in postmortem muscle tissue was developed and validated. Additionally, a validation of whole blood was carried out to compare the results from the two matrices. Solid-phase extraction was performed by an automated robotic system to minimize manual labour and risk of human errors, and increase robustness, sample throughput and sample traceability. The method was validated in terms of selectivity, matrix effect, extraction recovery, process efficiency, measuring range, lower limit of quantification, carry-over, stability, precision and accuracy. To correct for any inter-individual variability in matrix effects on analyte accuracy and precision, deuterated analogues of each analyte were used as internal standards. The lower limit of quantification in both blood and muscle homogenate ranged between 0.002 and 0.005 mg/kg, while the upper limit of quantification spanned from 0.20 to 1.0 mg/kg. Corrected with the 4-fold dilution factor, the corresponding concentrations in muscle tissue were 0.008-0.02 mg/kg at the lower limit of quantification and 0.80-4.0 mg/kg at the upper limit of quantification. The method showed acceptable precision and accuracy, with precision below 12% and accuracies ranging from 87% to 115% at up to 6 levels for all analytes in both matrices. In addition, comparison between calibration standards in spiked muscle homogenate and spiked blood showed that analyte concentrations in muscle samples could be quantified by using spiked blood samples as calibration standards with acceptable precision and accuracy when using deuterated analogues as internal standards. The investigation of matrix effects showed no great difference between blood and homogenates of non-decomposed and decomposed muscle tissue for most analytes. In the samples where high ion suppression or enhancement was observed, the results were corrected by the internal standards. Statistical comparison of quality control samples in blood and muscle tissue showed no obvious differences, and therefore muscle tissue was included in the routine method for analysis of blood samples and used in autopsy cases where no blood was available. By adding a semi-automated homogenization step before the remaining automated sample preparation, muscle tissue samples were easily incorporated into the workflow of the existing routine method. The present method has been successfully implemented in routine analysis of blood and muscle tissue since 2019., (Copyright © 2021 The Authors. Published by Elsevier B.V. All rights reserved.)

Purpose: In order to provide guidance and prepare ward managers for future crisis situations similar to the COVID-19 pandemic, the aim of this study was to reflect and learn how person-centred nursing leadership may be strengthened in such situations., Background: The pandemic has forced nurse leaders to face new challenges. Knowledge about their experiences may contribute to advancing leadership practices in times of future crises., Methods: A qualitative directed content analysis was chosen. The theoretical perspective was person-centred leadership. Thirteen ward managers from a Danish university hospital were included and interviewed using telephone interviews three months after the first national COVID-19 case was confirmed., Findings: The main findings of the study revealed that the ward managers often experienced a lack of timely, relevant information, involvement in decision-making and acknowledgement from the head nurse of department and the executive management. This was caused by the existing organizational cultures and the traditional hierarchy of communication. This meant that the ward managers' sense of own competences and leadership values and beliefs came under high pressure when they had to balance different stakeholders' needs., Conclusion: When the experience of ward managers results in them being unable to lead authentically and competently in a crisis like the COVID-19 pandemic, lack of engagement can occur, with serious consequences for patients, staff and the ward managers themselves. Traditional organizational cultures that are hierarchical and controlling needs to be challenged and reoriented towards collaborative, inclusive and participative practices of engagement and involvement. Leadership development must be an established and integrated component of organizations, so that ward managers are able to sustain person-centred ways of being and doing in times of crisis., Competing Interests: The authors report no conflicts of interest in this work., (© 2021 Hølge-Hazelton et al.)

The brainstem dorsal vagal complex (DVC) is known to regulate energy balance and is the target of appetite-suppressing hormones, such as glucagon-like peptide 1 (GLP-1). Here we provide a comprehensive genetic map of the DVC and identify neuronal populations that control feeding. Combining bulk and single-nucleus gene expression and chromatin profiling of DVC cells, we reveal 25 neuronal populations with unique transcriptional and chromatin accessibility landscapes and peptide receptor expression profiles. GLP-1 receptor (GLP-1R) agonist administration induces gene expression alterations specific to two distinct sets of Glp1r neurons-one population in the area postrema and one in the nucleus of the solitary tract that also expresses calcitonin receptor (Calcr). Transcripts and regions of accessible chromatin near obesity-associated genetic variants are enriched in the area postrema and the nucleus of the solitary tract neurons that express Glp1r and/or Calcr, and activating several of these neuronal populations decreases feeding in rodents. Thus, DVC neuronal populations associated with obesity predisposition suppress feeding and may represent therapeutic targets for obesity.

Background: Application of patient-reported outcome measures (PROM) seems to be a step toward person-centered care and identifying patients' unmet needs., Objective: To investigate the experiences of nurses when PROMs were introduced in a hematological clinical practice as part of a multimethod intervention study., Methods: The qualitative framework was guided by the interpretive description (ID) methodology, including a focused ethnographic approach with participant observations and interviews. The instruments introduced were the European Organization for Research and Treatment of Cancer Quality of Life Questionnaire Core 30 and the Outcomes and Experiences Questionnaire. Analysis was inspired by Habermas' critical theory., Results: The analysis revealed 2 predominant themes of nurses' experiences: "PROMs are only used when there is time-which there rarely is" and "PROMs cannot be used without a strategy, just because they are present.", Conclusions: Nurses' experiences with PROMs depended on the systems' rationale, resulting in limited capacity to use and explore PROMs. Nurses believed that PROMs might have the potential to support clinical practice, as PROMs added new information about patients' conditions but also identified needs within supportive care, leaving the potential of PROMs uncertain. Simply introducing PROMs to practice does not necessarily actuate their potential because use of PROMs is dependent on institutional conditions and mandatory tasks are prioritized., Implications for Practice: This study contributes knowledge of nurses' experiences when introducing PROMs in a hematological outpatient clinical practice. Findings can guide future PROMs research within the field of nursing., Competing Interests: The authors have no conflicts of interest to disclose., (Copyright © 2020 Wolters Kluwer Health, Inc. All rights reserved.)

Purpose: The aim was to identify the differences in experiences of Danish healthcare leaders in the beginnning of the coronavirus (COVID-19) pandemic and to generate knowledge for future leadership during and post crises., Background: The global spread of COVID-19 has affected healthcare systems worldwide and has forced healthcare leaders to face challenges few were prepared for. It is expected that the pandemic may hit in several waves within the next year and therefore healthcare leaders must be prepared for these waves., Methods: An online survey was developed, and comparative analyses were performed., Results: One hundred and sixty hospital leaders were invited, and 72% completed the questionnaire. Significant differences were found within three selected characteristics: 1) Management level: significantly more heads of departments experienced taking complex decisions ( P =0.05), being able to work in a way consistent with their beliefs and values ( P =0.05), and they were less likely to experience that collaboration with other leaders was adversely affected by the COVID-19 situation compared to ward managers ( P =0.04). On the other hand, ward managers were significantly more often worried about both their own health ( P =0.01) and their family's health ( P =0.04). 2) Management education: those with a formal management education more often experienced having the managerial competences to effectively manage the COVID-19 situation ( P =0.00), and performing meaningful tasks during the situation ( P =0.04). 3) Years of experience: significantly more leaders with more than five years of experience identified having the managerial competences to effectively manage the situation ( P =0.01)., Conclusion: Leadership support during a healthcare crisis like the COVID-19 pandemic should strategically focus on ward managers, leaders with no formal management education and leaders with less than two years of experience. Hospital leaders may use this knowledge to re-contextualize what is already known about targeted leadership support during healthcare crises and to act accordingly., Competing Interests: The authors report no conflicts of interest in this work., (© 2021 Hølge-Hazelton et al.)

Antimicrobial agents are used extensively off-label in mink, as almost no agents are registered for this animal species. Pharmacokinetic (PK) and pharmacodynamic (PD) data are required to determine antimicrobial dosages specifically targeting mink bacterial pathogens. The aims of this study were to assess, in a PKPD framework, the empirical dosage regimen for a combination of trimethoprim (TMP) and sulfadiazine (SDZ) in mink, and secondarily to produce data for future setting of clinical breakpoints. TMP and SDZ PK parameters were obtained experimentally in 22 minks following IV or oral administration of TMP/SDZ (30 mg/kg, i.e. 5 mg/kg TMP and 25 mg/kg SDZ). fAUC/MIC with a target value of 24 hr was selected as the PKPD index predictive of TMP/SDZ efficacy. Using a modeling approach, PKPD cutoffs for TMP and SDZ were determined as 0.062 and 16 mg/L, respectively. By incorporating an anticipated potentiation effect of SDZ on TMP against Escherichia coli and Staphylococcus delphini, the PKPD cutoff of TMP was revised to 0.312 mg/L, which is above the tentative epidemiological cutoffs (TECOFF) for these species. The current empirical TMP/SDZ dosage regimen (30 mg/kg, PO, once daily) therefore appears adequate for treatment of wild-type E. coli and S. delphini infections in mink., (© 2020 John Wiley & Sons Ltd.)

Background: PROMs can help healthcare professionals gain an improved understanding of patients' physical burdens, functional levels, and (health-related) quality of life throughout disease and medical treatment. The aim of this study was to investigate the barriers and potential opportunities PROMs may present in a haematological outpatient clinic from three different perspectives: patients, nurses and haematologists., Methods: The present study synthesizes three previously published studies that separately explored the experiences of patients, nurses and haematologists when implementing PROMs. The studies were all guided by the qualitative methodology Interpretive Description, including a focused ethnographic approach, to develop implications for future practice., Results: The overall themes that emerged from the analysis were "Structural similarities influence the adoption of PROMs" and "Different perspectives on the potential of PROMs.", Conclusion: Across the different user groups in the haematological outpatient clinic, the use of PROMs was thwarted due to an unquestioned commitment to biomedical knowledge and the system's rationality and norms: PROM data was not used in patient consultations. Nurses and haematologists expressed different preferences related to potential future PROMs and different objectives for PROMs in clinical practice. From the different perspectives of the patients, nurses and haematologists, PROMs were not compatible with clinical practice. Further research is recommended to develop PROMs validated for use in haematological outpatient clinics. Moreover, implementation strategies adjusted to the structural barriers of the system are crucial.

Background: Patient-reported outcome measures (PROMs) in clinical practice have the potential to contribute to and support shared decision-making processes by giving voice to patient concerns during consultations. However, the perspectives of patients diagnosed with chronic hematologic cancer on the use of PROMs are unknown., Objective: To describe how patients diagnosed with hematologic cancer experience participating in a randomized PROM intervention study, including initial invitation, completion of questionnaires, and outpatient clinic visits., Methods: A qualitative conceptual framework guided the study, using Interpretive Description with a focused ethnographic approach to explore patient experiences with PROMs in applied practice. Analysis was inspired by Habermas' social theory of communicative action., Results: The analysis revealed 3 predominant themes of patient experiences: that PROMs were "In the service of a good cause," "The questions are not really spot on," and "PROMs are sometimes used for something," that is, unknown to the patient., Conclusions: The patients' experiences were dominated by the perspective of the healthcare system and by gratitude and imbalanced power relations. During completion of questionnaires, patients struggled to identify with items, and the questionnaires were associated with low content validity. When visiting the outpatient clinic, patients reported that doctors and nurses rarely discussed patients' PROMs., Implications for Practice: This study contributes knowledge of patient experiences of the integration of PROMs in hematologic outpatient clinical practice. Findings can guide further research and improve future implementation of PROMs.

The kinetic theory of enzymes that modify insoluble substrates is still underdeveloped, despite the prevalence of this type of reaction both in vivo and industrial applications. Here, we present a steady-state kinetic approach to investigate inhibition occurring at the solid-liquid interface. We propose to conduct experiments under enzyme excess (E0 ≫ S0), i.e. the opposite limit compared with the conventional Michaelis-Menten framework. This inverse condition is practical for insoluble substrates and elucidates how the inhibitor reduces enzyme activity through binding to the substrate. We claim that this type of inhibition is common for interfacial enzyme reactions because substrate accessibility is low, and we show that it can be analyzed by experiments and rate equations that are analogous to the conventional approach, except that the roles of enzyme and substrate have been swapped. To illustrate the approach, we investigated the major cellulases from Trichoderma reesei (Cel6A and Cel7A) acting on insoluble cellulose. As model inhibitors, we used catalytically inactive variants of Cel6A and Cel7A. We made so-called inverse Michaelis-Menten curves at different concentrations of inhibitors and found that a new rate equation accounted well for the data. In most cases, we found a mixed type of surface-site inhibition mechanism, and this probably reflected that the inhibitor both competed with the enzyme for the productive binding-sites (competitive inhibition) and hampered the processive movement on the surface (uncompetitive inhibition). These results give new insights into the complex interplay of Cel7A and Cel6A on cellulose and the approach may be applicable to other heterogeneous enzyme reactions., (© 2020 The Author(s). Published by Portland Press Limited on behalf of the Biochemical Society.)

In forensic investigations, such as drug-facilitated crimes, reference values are useful for interpretation of hair results. The aim of this study was to establish levels of zopiclone and two main metabolites, N-desmethylzopiclone and zopiclone N-oxide, in hair after the administration of a single dose of zopiclone, as very limited data are published. A controlled study was performed, where 16 volunteers consumed either 5 or 10mg zopiclone. Hair was sampled prior to consumption and 14, 30, 60, and 120 days after intake. The deposition of drug in hair segments of all sampling time points was followed in small hair segments of 5-mm, using a validated ultra-high performance liquid chromatography-tandem mass spectrometry method. In all participants, hair segments corresponding to the time of intake were positive for zopiclone, but also with lower concentrations in the neighbouring segments. The highest zopiclone concentrations were detected in samples collected 30 or 60 days after intake. For all sampling time points maximum values for the 5-mg dose ranged from 5.0-370pg/mg for zopiclone and 5.4 to 300pg/mg for N-desmethylzopiclone, where the maximum values for the 10-mg dose ranged from 17 to 590pg/mg for zopiclone and 25-410pg/mg for N-desmethylzopiclone for all sampling time points. No significant difference in concentrations was found between the two dosing groups for either zopiclone or N-desmethylzopiclone. Almost half of the participants showed lower levels 14 days after intake than in the later sampling time points. The metabolite to parent drug ratio of N-desmethylzopiclone to zopiclone varied from 0.6 to 3.4 (median=1.2) for the maximum levels of all sampling time points. N-desmethylzopiclone are suggested to serve as an additional marker to confirm the intake of zopiclone. Traces of zopiclone N-oxide were detected in hair from only eight participants. This study showed, that it was possible to follow zopiclone and N-desmethylzopiclone in hair for 4 months even though the drugs was divided into several segments in the latest collected hair samples, and no obvious wash-out effect between the sampling time points by e.g. personal hygiene could be discerned because the cumulated amount at each sampling time point was similar. We conclude that the analysis of short segments e.g. segments of 5-mm can help determine the time of a single intake of zopiclone and that obtaining a sample 1-2 months after a drug exposure provide the best conditions to detect and interpret the results., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Background: The patient-doctor relationship is crucial to provide person-centred care, allowing the alleviation of symptom burden caused by disease or treatment. Implementing Patient Reported Outcome Measures (PROMs) is suggested to inform the decision-making process and lead to initiation of care. Yet there are knowledge gaps regarding how meaningful it is to incorporate PROMs in clinical settings. The aim of this study was to investigate haematologists' experiences when PROMs were implemented in an outpatient setting., Methods: Fourteen participant observations, 13 individual interviews and three in-depth interviews were conducted with haematologists, guided by the qualitative methodology Interpretive Description. Analysis was inspired by Habermas' critical theoretical framework., Results: The haematologists included were characterised by dichotomous experiences with PROMs, either resistant to or supporting their implementation. None were observed to elaborate on PROMs during consultations: instead, primary attention was spent discussing the hematological agenda dictated by the system., Conclusion: The use of PROMs for individualized care was linked with extensive uncertainties and PROMs were not requested by the haematologists. To improve individualized care, other approaches may be more suitable. If PROMs are to be incorporated into future clinical practice, they should be tested tothe specific patient group and involve relevant users.

Purpose: Eir version 3 (V3) is an electronic tool for administration of patient-reported outcome measures (Eir-Patient) that immediately presents patient scores on the physician's computer (Eir-Doctor). Perceived usability is an important determinant for successful implementation. The aim of this study was to answer the following research question evaluated at the cancer outpatient clinics, in the patients' home, and at general practitioners' (GPs) offices: What are the number, type, and severity of usability issues evaluated by the patient (Eir-Patient module) and by the physician (Eir-Doctor module)?, Methods: A usability evaluation using observations, think-aloud sessions, individual interviews and focus group interviews in cancer patients and their physicians was conducted. Identified usability issues were graded on a severity scale from 1 (irritant) to 4 (unusable)., Results: Overall, 73 Eir registrations were performed by 37 patients, and used by 17 physicians in clinical consultations. All patients were able to complete the Eir-Patient symptom registration. Seventy-two usability issues were identified. None of them were graded as unusable. For the Eir-Patient module, 62% of the identified usability issues was graded as irritant (grade 1), 18% as moderate (grade 2), and 20% as severe (grade 3). For the Eir-Doctor module, 46% of the identified usability issues were graded as irritant, 36% as moderate and 18% as severe., Conclusions: In the updated Eir version, issues in the severe and moderate categories have been changed, to optimize the usability of using real-time PROMs in clinical practice.

Studies of drug permeability rate and localization in buccal mucosa are essential to gain new knowledge of means such as chemical enhancers or osmolality to enhance buccal drug transport in the development of new buccal drug products. The transport of caffeine, diazepam and mannitol across porcine buccal mucosa was studied in modified Ussing chambers with a hypotonic donor solution, in the presence of levulinic acid (LA), oleic acid (OA), propylene glycol (PG) as well as sodium dodecyl sulfate (SDS). Subsequently, matrix-assisted laser desorption ionization - mass spectrometry imaging (MALDI-MSI) was applied to image the spatial distribution of caffeine, mannitol and SDS in cross-sections of porcine buccal mucosa. The results revealed that none of the permeation enhancing strategies improved the permeability of caffeine or diazepam, despite impact on the tissue integrity by OA and SDS, as seen by an increased permeability of mannitol. Further studies are needed with OA since PG solvent may have concealed the possible impact of OA. SDS decreased the permeability of caffeine and diazepam, a decrease which can be explained by micellar lipid extraction and encapsulation in micelles. MALDI-MSI showed that SDS permeated into approximately one-third of the epithelium, and it therefore appears that the main permeability barrier for mannitol is located in the outer epithelium. MALDI-MSI was shown to be a useful method for imaging spatial distribution of drugs and permeations enhancers in buccal mucosa., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Vitamin C (vitC) is important in the developing brain, acting both as an essential antioxidant and as co-factor in the synthesis and metabolism of monoaminergic neurotransmitters. In guinea pigs, vitC deficiency results in increased oxidative stress, reduced hippocampal volume and neuronal numbers, and deficits in spatial memory. This study investigated the effects of 8 weeks of either sufficient (923 mg vitC/kg feed) or deficient (100 mg vitC/kg feed) levels of dietary vitC on hippocampal monoaminergic neurotransmitters and markers of synapse formation in young guinea pigs with spatial memory deficits. Western blotting and high performance liquid chromatography (HPLC) were used to quantify the selected markers. VitC deficiency resulted in significantly reduced protein levels of synaptophysin ( p = 0.016) and a decrease in 5-hydroxyindoleacetic acid/5-hydroxytryptamine ratio ( p = 0.0093). Protein expression of the N -methyl-d-aspartate receptor subunit 1 and monoamine oxidase A were reduced, albeit not reaching statistical significance ( p = 0.0898 and p = 0.067, respectively). Our findings suggest that vitC deficiency induced spatial memory deficits might be mediated by impairments in neurotransmission and synaptic development.

Approximately 15% of the Western world population, including pregnant women and their children, is characterized as vitamin C (vitC) deficient. In guinea pigs, early life vitC deficiency causes spatial memory deficits, decreased hippocampal volume and neuron numbers, in otherwise clinically healthy animals. We hypothesized that vitC deficiency leads to decreased brain-derived neurotrophic factor and synaptic plasticity markers in selected brain areas (frontal cortex, hippocampus and striatum) and cause morphological changes in cornu ammonis 1 pyramidal neurons of the hippocampus either through a direct effect or indirectly by increased oxidative stress. Fifty-seven female guinea pigs were allocated to three groups receiving either 1390, 100 or 0⁻50 mg vitC/kg feed for 11 weeks. Dietary vitC levels were reflected in the plasma, cortical and adrenal gland levels, however, redox imbalance was only present in the adrenal glands allowing for the investigation of a direct influence of vitC deficiency on the chosen parameters in the brain. Synaptic plasticity markers were not affected in the investigated brain areas and no differences in isolated pyramidal neuron morphology was recorded. Based on our findings, it appears that vitC deficiency may primarily elicit impaired neuronal function through increased levels of oxidative stress.